Human pegylated interleukin 22 Y51A antagonist - CYT-20 GENE ID 50616

Mono-pegylated (with 20 kDa PEG)recombinant human interleukin 22 Y51A mutant (numbering according to human IL-22 including signal peptide) is one polypeptide chain containing 147 amino, an additional Ala at N-terminus and one molecule of PEG 20 kDa at its N-terminus, having an expected molecular mass of ~ 36 kDa as determined by MS. However due to enlarged hydrodymanic volume it runs on the SGS-Page as 50 kDa protein and in gel-filtration on Superdex 200 as over 200 kDa protein. It was prepared similarly to that described by L. Niv-Spector et al. Protein Eng Des Sel. (PEDS) 25:397-404 (2012).

E. coli

 

 

White lyophilized powder

The protein was lyophilized from a concentrated (1.0 mg/ml) solution with 0.003mM NaHCO3.

It is recommended to reconstitute the lyophilized pegylated mouse IL22 Y51A in sterile water or sterile 0.4% NaHCO3 adjusted to pH 8-9, not less than 100µg/ml, which can then be further diluted with other aqueous solutions.

Lyophilized pegylated mouse IL22 although stable at room temperature for several days, should be stored desiccated below -20C. Upon reconstitution at > 0.1 pegylated mIL22 mg/ml and up to 2 mg/ml and filter sterilization it can be stored at 4C or even room temperature for several weeks making it suitable for long term infusion studies using osmotic pumps. At lower concentration addition of a carrier protein (0.1% HSA or BSA) is suggested. Please prevent freeze-thaw cycles.

Greater than 98.0% as determined by:
(a) Gel filtration analysis.
(b) Analysis by reducing and non-reducing SDS-PAGE gel.

The sequence of the N-terminal amino acids could not be determined indicating that the PEG moiety is attached to the N-terminus.

The purified mouse pegylated IL22 consists of > 90% monomers as determined by gel-filtration chromatography.

PLR’s human IL-22 Y51A mutant is biologically active, capable of full inhibition of mouse IL-22 induced STAT3 phosphorylation in HepG cells. Its inhibitory acrtivity in vitro is ~ 10-20% compared to the non-pegylated mIL22 (Y51A) mutant.

Less than 0.05 ng/µg (0.5 EU/µg) of recombinant pegylated IL-22 Y51A.

Protein quantitation of human interleukin 22 Y51A mutant was carried out by UV spectroscopy at 280 nm using the absorbency value of 0.18 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the DNAman computer analysis program of protein sequences.

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